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    products/primary-antibodies/vav3-antibody-ep1130y-ab52938.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-VAV3 antibody [EP1130Y] (ab52938)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (6)

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Western blot - Anti-VAV3 antibody [EP1130Y] (ab52938)
  • Western blot - Anti-VAV3 antibody [EP1130Y] (ab52938)
  • Immunocytochemistry/ Immunofluorescence - Anti-VAV3 antibody [EP1130Y] (ab52938)
  • Immunoprecipitation - Anti-VAV3 antibody [EP1130Y] (ab52938)
  • Flow Cytometry (Intracellular) - Anti-VAV3 antibody [EP1130Y] (ab52938)
  • Anti-VAV3 antibody [EP1130Y] (ab52938)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1130Y] to VAV3
  • Suitable for: WB, Flow Cyt (Intra), IP, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human VAV3 knockout HeLa cell line (ab265419)

View more associated products

Overview

  • Product name

    Anti-VAV3 antibody [EP1130Y]
    See all VAV3 primary antibodies
  • Description

    Rabbit monoclonal [EP1130Y] to VAV3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt (Intra), IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Jurkat and HeLa cell lysates. ICC/IF: Jurkat cells; IP: Jurkat whole cell lysate; Flow Cyt (intra): Jurkat cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1130Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • Small G Proteins
    • Regulators
    • Cancer
    • Cell cycle
    • Cell division
    • Cancer
    • Signal transduction
    • G protein signaling
    • Small G proteins
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-VAV3 antibody [EP1130Y] (ab206236)
    • Anti-VAV3 antibody [EP1130Y] - BSA and Azide free (ab247340)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
  • Substrate reagent

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52938 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 98 kDa).
Flow Cyt (Intra)
1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP
1/30.
ICC/IF
1/50.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 98 kDa).
Flow Cyt (Intra)
1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP
1/30.
ICC/IF
1/50.

Target

  • Function

    Exchange factor for GTP-binding proteins RhoA, RhoG and, to a lesser extent, Rac1. Binds physically to the nucleotide-free states of those GTPases. Plays an important role in angiogenesis. Its recruitement by phosphorylated EPHA2 is critical for EFNA1-induced RAC1 GTPase activation and vascular endothelial cell migration and assembly.
  • Sequence similarities

    Contains 1 CH (calponin-homology) domain.
    Contains 1 DH (DBL-homology) domain.
    Contains 1 PH domain.
    Contains 1 phorbol-ester/DAG-type zinc finger.
    Contains 1 SH2 domain.
    Contains 2 SH3 domains.
  • Target information above from: UniProt accession Q9UKW4 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10451 Human
    • Omim: 605541 Human
    • SwissProt: Q9UKW4 Human
    • Unigene: 267659 Human
    • Alternative names

      • FLJ40431 antibody
      • Guanine nucleotide exchange factor VAV3 antibody
      • Protein vav 3 antibody
      • Protein vav3 antibody
      • RGD1565941 antibody
      • VAV 3 antibody
      • Vav 3 guanine nucleotide exchange factor antibody
      • VAV 3 oncogene antibody
      • VAV 3 protein antibody
      • VAV-3 antibody
      • Vav3 antibody
      • VAV3 oncogene antibody
      • VAV3 protein antibody
      • VAV3_HUMAN antibody
      see all

    Images

    • Western blot - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Western blot - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Lanes 1 & 3 : Anti-VAV3 antibody [EP1130Y] (ab52938) at 1/1000 dilution


      Lane 1 : Wild-type HeLa cell lysate at 20 µg
      Lane 2 : Empty
      Lane 3 : VAV3 knockout HeLa cell lysate at 20 µg

      Performed under reducing conditions.

      Predicted band size: 98 kDa
      Observed band size: 98 kDa



      False colour image of Western blot: Anti-VAV3 antibody [EP1130Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab52938 was shown to bind specifically to VAV3. A band was observed at 98 kDa in wild-type HeLa cell lysates with no signal observed at this size in VAV3 knockout cell line ab265419 (knockout cell lysate ab258280). To generate this image, wild-type and VAV3 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Western blot - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Anti-VAV3 antibody [EP1130Y] (ab52938) at 1/1000 dilution (Purified) + Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 98 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Immunocytochemistry/ Immunofluorescence - Anti-VAV3 antibody [EP1130Y] (ab52938)

      Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling VAV3 with purified ab52938 at 1/50 dilution (7.2 µg/mL). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunoprecipitation - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Immunoprecipitation - Anti-VAV3 antibody [EP1130Y] (ab52938)

      Purified ab52938 at 1/30 dilution (2µg) immunoprecipitating VAV3 in Jurkat whole cell lysate.
      Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg.
      Lane 2 (+): ab52938 + Jurkat whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52938 in Jurkat whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-VAV3 antibody [EP1130Y] (ab52938)
      Flow Cytometry (Intracellular) - Anti-VAV3 antibody [EP1130Y] (ab52938)

      Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling VAV3 with purified ab52938 at 1/40 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Anti-VAV3 antibody [EP1130Y] (ab52938)
      Anti-VAV3 antibody [EP1130Y] (ab52938)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (6)

    Publishing research using ab52938? Please let us know so that we can cite the reference in this datasheet.

    ab52938 has been referenced in 6 publications.

    • Fray MA  et al. Vav2 lacks Ca2+ entry-promoting scaffolding functions unique to Vav1 and inhibits T cell activation via Cdc42. J Cell Sci 133:N/A (2020). PubMed: 31974114
    • Xiao Y  et al. LINC00265 targets miR-382-5p to regulate SAT1, VAV3 and angiogenesis in osteosarcoma. Aging (Albany NY) 12:20212-20225 (2020). PubMed: 33109774
    • Maruyama K  et al. Bone-protective Functions of Netrin 1 Protein. J Biol Chem 291:23854-23868 (2016). PubMed: 27681594
    • Aguilar H  et al. VAV3 mediates resistance to breast cancer endocrine therapy. Breast Cancer Res 16:R53 (2014). Mouse . PubMed: 24886537
    • Nomura T  et al. Targeting the Vav3 oncogene enhances docetaxel-induced apoptosis through the inhibition of androgen receptor phosphorylation in LNCaP prostate cancer cells under chronic hypoxia. Mol Cancer 12:27 (2013). ICC/IF ; Human . PubMed: 23566222
    • Yi F  et al. Mechanism of homocysteine-induced Rac1/NADPH oxidase activation in mesangial cells: role of guanine nucleotide exchange factor Vav2. Cell Physiol Biochem 20:909-18 (2007). WB . PubMed: 17982273

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question

    I loaded 20 ug of whole cell lysate and ran an SDS-PAGE. After transfer, the PVDF was blocked for 1 hour in milk and then incubated with primary antibody. The first antibody that I purchased was tried at various dilutions from 1:300 to 1:10,000, all in milk for one hour at room temperature. Secondary antibody was 1:10,000 for one hour at room temperature. I have tried the second antibody at 1:500 and 1:1000, both in milk. There are >10 bands on a 10% gel on the shortest exposure. I have tried both Hela cells and a bronchial epithelial cell line. The dominant bands are doublets at 56 and 34 kDa. I transfected these cells with siRNAs to Vav3 and see no decrease in levels in any of the bands compared to control with the exception of one band at ˜32 kDa. Any explanation for these results? Thanks for your help

    Read More

    Abcam community

    Verified customer

    Asked on Jan 03 2012

    Answer

    Thanks for the reply. I am a little stumped as to why you are having these issues. I can only think of 2 things that may help (and I am sure you have already tried them): 1 - One reason for multiply bands is that there is not enough protease inhibitor in your sample buffer. Increasing the concentration of this would help prevent breakdown of your protein. 2 - Switching to a 5% BSA block would also help reduce the non-specific binding that is occurring. Other than that, I honestly cannot think what else you could try. If you have already tried the above variations and they did not prove successful then I would be happy to either replace this antibody or refund the original purchase. I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Jan 03 2012

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